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Cerebral ischemia triggers a cascade of events that contribute to ischemic brain damages. Zinc release and accumulation has been shown to lead to brain cell death following cerebral ischemia. However, the mechanism underlying remains to be elucidated. Our recently published work showed that suppression of mitochondrial-derived reactive oxygen species (ROS) production significantly reduced ischemic stroke related brain damage within 6 h. Herein, we investigated the relationship between zinc accumulation and mitochondrial-derived ROS production in astrocytes after 3-h hypoxia. We found that inhibition of mitochondrial-derived ROS significantly decreased total amount of ROS generation and cell death in primary astrocytes during hypoxia when zinc was overload. In contrast, the inhibition of NADPH oxidase-derived ROS had less of an effect. Our results also showed that zinc and mitochondria were colocalized in hypoxic astrocytes. Moreover, extracellular zinc addition caused zinc accumulation in the mitochondria and decreased mitochondrial membrane potential, leading to mitochondria dysfunction. These findings provide a novel mechanism that zinc accumulation contributes to hypoxia-induced astrocytes death by disrupting mitochondria function, following cerebral ischemia.
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